CN107050507A - A kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material - Google Patents
A kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material Download PDFInfo
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- CN107050507A CN107050507A CN201710330841.2A CN201710330841A CN107050507A CN 107050507 A CN107050507 A CN 107050507A CN 201710330841 A CN201710330841 A CN 201710330841A CN 107050507 A CN107050507 A CN 107050507A
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- poly phosphazene
- dopa
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- structural modification
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
- A61L27/34—Macromolecular materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
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Abstract
A kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material, belongs to composite biological material preparation field, and transformation of the way Preparation Method mainly includes：Prepare the amine-modified solution of DOPA, prepare poly phosphazene static spinning membrane, the spraying amine-modified solution of DOPA.By DOPA structural modification poly phosphazene tissue renovation material made from this method, the cell adhesion and bioactivity of tissue renovation material are further increased.
Description
Technical field
The invention belongs to composite biological material preparation field, and in particular to a kind of DOPA structural modification poly phosphazene tissue repair
The preparation method of material.
Background technology
Biomaterial, which is developed so far, has gone through three Main Stages.Since 1980s, with medical treatment, health care
And the bio-medical material promoted for the purpose of quality of life etc. achieves quick development, has had more than 50 kinds of implantation instrument quilts
Applied to clinic, the engineering material of technology maturation is typically derived from, and with biocompatibility and defective tissue alternative functions,
Such as heart artificial blood vessel, joint prosthesis and the artificial kidney for being widely used to clinic.Above-mentioned bio-medical material, with one
Universal general character:Biologically inert.I.e. bio-medical material develops followed principle and is to try to acceptor to the different of implantation instrument
Thing reaction is preferably minimized.From last century eighties the to the nineties, the emphasis of biomedical materials field is gradually turned by biologically inert
To bioactivity and controlled degradation, second generation bio-medical material and product are developed.
Later stage 1990s, third generation biomaterial is accompanied by the foundation of organizational engineering, develops and send out rapidly
What exhibition was got up.Using Tissue Engineering Biomaterials support as representative, it combines engineering science and life science principle, careful to plant
Born of the same parents, which provide, is adapted to its growth, matrix synthesis and the biology space for playing other functions, overcomes in the past single cell and moves
The shortcomings of cell is difficult to survive in plant, matrix synthesis capability is low, the structure for engineered tissue provides cell carrier
And structure stand.By bioactive materials and degradation material, the two independent concepts have been combined this kind of bio-medical material
Come, molecular modification is carried out in degradation material, the interaction for causing cellular integration plain, induced cell proliferation, differentiation, and
The synthesis and assembling of extracellular matrix.But the performance for the biomaterial for being applied to organizational project at present need further to carry
It is high, it is impossible to meet the demand of modern medicine.
The content of the invention
In order to solve problems of the prior art, repaiied the invention provides a kind of DOPA structural modification poly phosphazene tissue
The preparation method of multiple material, further increases the cell adhesion and bioactivity of tissue renovation material.
The present invention uses following technical scheme：
A kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material, comprises the following steps：
Step one：By 350mg Gelatins in 60mL water, heating water bath is to 50 DEG C of stirrings to being completely dissolved obtained gelatin
Solution；
Step 2：By gelatin solution and 150mg N- hydroxysuccinimides, 191mg 1- ethyls-(3- dimethylaminos
Propyl group) phosphinylidyne diimine and 103mg dopamines mixing, in electric heating constant temperature tank 40 DEG C stirring 48 hours after, solution is placed in
In bag filter, it is put into the beaker for filling ultra-pure water, is placed in electric heating constant temperature tank and 2-3day is stirred under the conditions of 40 DEG C, and often
A ultra-pure water is changed every 8h；
Step 3：The solution remained in bag filter is taken out, is placed in -20 DEG C of household freezers and freezes 8h, then be placed on freeze dryer
48h is dried, adhesiveness gelatin is obtained；
Step 4：Adhesiveness gelatin is made to the 3mg/ml aqueous solution, pH value of water solution is adjusted to 7 with HCl and NaOH, i.e.,
Obtain the amine-modified solution of DOPA；
Step 5：Poly phosphazene is dissolved into trifluoroethanol, compound concentration is to stir under 45w/v%, normal temperature in favor of poly-
Phosphine nitrile is fully dissolved, and uniform spinning solution is obtained after 24h, and poly phosphazene static spinning membrane is obtained through electrostatic spinning, is placed in 50 DEG C very
In empty drying box, dry to constant weight；
Step 6：By the amine-modified solution even application of DOPA in static spinning membrane surface, and 12h is dried in 37 DEG C of drying boxes
Afterwards, rinsed 3 times with phosphate buffer (PBS), produce tissue renovation material.
It is preferred that, the bag filter remaining molecules amount described in step 2 is 10000Da.
It is preferred that, ultrapure water consumption is to cover bag filter in described step two.
It is preferred that, the spinning parameter of electrostatic spinning is in step 5：Copper mesh reception, voltage 12kv, reception are apart from 15cm, stream
Fast 0.4ml/h.
It is preferred that, the thickness sprayed in step 6 is 5-30nm.
The beneficial effects of the present invention are：
1) dopamine is a kind of compound in neurohormone, containing abundant ethylamino and catechol active function groups,
The surface of any body can be almost sticked to, particularly with organic surface effect more preferably.Gelatin after DOPA is amine-modified sticks
Property is higher, and forms on material surface dopamine-gelatin layer of polymerization.The modification of dopamine, is not only increased fixed
The content of gelatin, while promoting the adhesion of cell, strengthens the cell compatibility of material surface.
2) substantial amounts of hydroxyl and a small amount of carboxyl and amino are contained in gelatin molecule structure, with extremely strong hydrophily.Cause
This, with the amino of dopamine condensation reaction can occur for the carboxyl in gelatin molecule.There are gelatin other synthetic materials can not compare
Biocompatibility, degradability and the bioactivity of plan.
3) poly phosphazene has good biodegradability, while also having good Bioabsorbable and bio-compatible
Property, any environmental issue will not be left after degradation, and in medical field, oneself is considered as most promising degradable macromolecule material
Material.
Embodiment
The present invention is described in further detail with reference to embodiments.
Embodiment 1
A kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material, comprises the following steps：
Step one：By 350mg Gelatins in 60mL water, heating water bath is to 50 DEG C of stirrings to being completely dissolved obtained gelatin
Solution；
Step 2：By gelatin solution and 150mg N- hydroxysuccinimides, 191mg 1- ethyls-(3- dimethylaminos
Propyl group) phosphinylidyne diimine and 103mg dopamines mixing, in electric heating constant temperature tank 40 DEG C stirring 48 hours after, solution is placed in
In bag filter, described bag filter remaining molecules amount is 10000Da, and bag filter is put into the beaker for filling ultra-pure water, ultrapure
Beaker is placed in electric heating constant temperature tank and stirs 2-3day under the conditions of 40 DEG C by water consumption to cover bag filter position, and every
8h changes a ultra-pure water；
Step 3：The solution remained in bag filter is taken out, is placed in -20 DEG C of household freezers and freezes 8h, then be placed on freeze dryer
48h is dried, adhesiveness gelatin is obtained；
Step 4：Adhesiveness gelatin is made to the 3mg/ml aqueous solution, pH value of water solution is adjusted to 7 with HCl and NaOH, i.e.,
Obtain the amine-modified solution of DOPA；
Step 5：Poly phosphazene is dissolved into trifluoroethanol, compound concentration is to stir under 45w/v%, normal temperature in favor of poly-
Phosphine nitrile is fully dissolved, and uniform spinning solution is obtained after 24h, and poly phosphazene static spinning membrane is obtained through electrostatic spinning, is placed in 50 DEG C very
In empty drying box, dry to constant weight；
Step 6：By the amine-modified solution even application of DOPA in static spinning membrane surface, the thickness of spraying is 5-30nm, and
Dried in 37 DEG C of drying boxes after 12h, rinsed 3 times with phosphate buffer (PBS), produce tissue renovation material.
Cell adhesion experiments：The HUVEC cells of culture are taken, are digested, cell count, with 5 × 103Cell/mL density is inoculated with
On tissue renovation material made from embodiment 1 and blank control group poly phosphazene material, per hole 1mL, after inoculation 0.5h, 1h, 2h,
Rinsed with sterile PBS solution, as a result Microscopic observation and counting of taking a picture see the table below 1.It can be seen that after DOPA is amine-modified
The cell quantity of the blank control group of the cell quantity that poly phosphazene tissue renovation material is adhered to significantly more than not by modification.
Table 1
Cell proliferation experiment：The HUVEC cells of culture are taken, are digested, cell count, with 5 × 103Cell/mL density is inoculated with
On tissue renovation material made from embodiment 1 and blank control group poly phosphazene material, per hole 1mL, liquid was changed every two days, is inoculated with
After 5day, Cell counting Kit WST-8 detects the cytoactive of material surface, the results are shown in Table 2, it can be deduced that pass through dopamine
The cytoactive of poly phosphazene tissue renovation material after modification is slightly above the cytoactive of blank assay group.
Table 2
Claims (5)
1. a kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material, it is characterised in that comprise the following steps：
Step one：By 350mg Gelatins in 60mL water, heating water bath is molten to obtained gelatin is completely dissolved to 50 DEG C of stirrings
Liquid；
Step 2：By gelatin solution and 150mg N- hydroxysuccinimides, 191mg 1- ethyls-(3- dimethylaminopropyls)
Phosphinylidyne diimine and 103mg dopamines mixing, in electric heating constant temperature tank 40 DEG C stirring 48 hours after, solution is placed in bag filter
In, it is put into the beaker for filling ultra-pure water, is placed in electric heating constant temperature tank and 2-3day is stirred under the conditions of 40 DEG C, and changed every 8h
Ultra-pure water；
Step 3：The solution remained in bag filter is taken out, is placed in -20 DEG C of household freezers and freezes 8h, then is placed on freeze dryer dry
48h, obtains adhesiveness gelatin；
Step 4：Adhesiveness gelatin is made to the 3mg/ml aqueous solution, pH value of water solution is adjusted to 7 with HCl and NaOH, i.e., much
The amine-modified solution of bar；
Step 5：Poly phosphazene is dissolved into trifluoroethanol, compound concentration is to stir in favor of poly phosphazene under 45w/v%, normal temperature
Uniform spinning solution is obtained after fully dissolving, 24h, poly phosphazene static spinning membrane is obtained through electrostatic spinning, 50 DEG C of vacuum is placed in and does
In dry case, dry to constant weight；
Step 6：By the amine-modified solution even application of DOPA in static spinning membrane surface, and dried in 37 DEG C of drying boxes after 12h,
Rinsed 3 times with phosphate buffer (PBS), produce tissue renovation material.
2. a kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material according to claim 1, its feature
It is：Bag filter remaining molecules amount described in step 2 is 10000Da.
3. a kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material according to claim 1, its feature
It is：Ultrapure water consumption is to cover bag filter in described step two.
4. a kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material according to claim 1, its feature
It is, the spinning parameter of electrostatic spinning is in step 5：Copper mesh reception, voltage 12kv, reception are apart from 15cm, flow velocity 0.4ml/h.
5. a kind of preparation method of DOPA structural modification poly phosphazene tissue renovation material according to claim 1, its feature
It is：The thickness sprayed in step 6 is 5-30nm.
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| Application Number | Priority Date | Filing Date | Title |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101257935A (en) * | 2005-09-05 | 2008-09-03 | 百润生物技术公司 | Multi-layered antiadhesion barrier |
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2017
- 2017-05-11 CN CN201710330841.2A patent/CN107050507A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101257935A (en) * | 2005-09-05 | 2008-09-03 | 百润生物技术公司 | Multi-layered antiadhesion barrier |
Non-Patent Citations (1)
| Title |
|---|
| 杨茜: ""多巴结构用于材料表面的仿生修饰及其生物学基础研究"", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
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